Altered lipid composition and enhanced lipid production in green microalga by introduction of brassica diacylglycerol acyltransferase 2

Higher lipid biosynthesis and accumulation are important to achieve economic viability of biofuel production via microalgae. To enhance lipid content, Chlamydomonas reinhardtii was genetically engineered with a key enzyme diacylglycerol acyltransferase (BnDGAT2) from Brassica napus, responsible for neutral lipid biosynthesis. The transformed colonies harbouring aph7 gene, screened on hygromycin‐supplemented medium, achieved transformation frequency of ~120 ± 10 colonies/1 × 10⁶ cells. Transgene integration and expression were confirmed by PCR, Southern blots, staining lipid droplets, proteins and spectro‐fluorometric analysis of Nile red‐stained cells. The neutral lipid is a major class (over 80% of total lipids) and most significant requirement for biodiesel production; this was remarkably higher in the transformed alga than the untransformed control. The levels of saturated fatty acids in the transformed alga decreased to about 7% while unsaturated fatty acids increased proportionately when compared to wild type cells. Polyunsaturated fatty acids, especially α‐linolenic acid, an essential omega‐3 fatty acid, were enhanced up to 12% in the transformed line. Nile red staining confirmed formation of a large number of lipid globules in the transformed alga. Evaluation of long‐term stability and vitality of the transgenic alga revealed that cryopreservation produced significantly higher quantity of lipid than those maintained continuously over 128 generations on solid medium. The overexpression of BnDGAT2 significantly altered the fatty acids profile in the transformed alga. Results of this study offer a valuable strategy of genetic manipulation for enhancing polyunsaturated fatty acids and neutral lipids for biofuel production in algae.     

UV protection in the photosymbiotic ascidian Didemnum molle inhabiting different depths

Didemnum molle is a colonial ascidian that harbors the prokaryotic photosymbiont Prochloron in its cloacal cavity. Colonies occur over a relatively wide bathymetric range (approximately 0-30 m), and colony color is widely variable, partly depending on depth. Colonies in shallow sites are bright white, with densely distributed spicules, and often with brown or dark gray pigmentation, while colonies in deeper sites are less pigmented, with sparsely distributed spicules. Didemnum molle colonies contain mycosporine-like amino acids (MAAs) as UV-absorbing substances. These include mycosporine-glycine, shinorine, and porphyra-334. Among colonies from 5-, 10-, 15-, and 20-m depths, the concentration of total MAAs was significantly high at 10 m and low at 20 m. Colonies at 10 m need to maintain low spicule densities to have enough photosynthetically active radiation (PAR) to maintain the photosymbionts, and they probably concentrate MAAs to block UV radiation without attenuating PAR. Because high levels of PAR cause photoinhibition of photosynthesis, spicules and pigment cells would be more effective for photoprotection in shallow water. Colonies of D. molle may adjust the light conditions for photosymbionts by combining MAAs, spicules, and pigment cells in varying amounts.     

Natural ultraviolet radiation and photosynthetically active radiation induce formation of mycosporine-like amino acids in the marine macroalga Chondrus crispus (Rhodophyta)

The UV-absorbing mycosporine-like amino acids (MAAs) are hypothesized to protect organisms against harmful UV radiation (UVR). Since the physiology and metabolism of these compounds are unknown, the induction and kinetics of MAA biosynthesis by various natural radiation conditions were investigated in the marine red alga Chondrus crispus collected from Helgoland, Germany. Three photosynthetically active radiation (PAR, 400–700 nm) treatments without UVR and three UV-A/B (290–400 nm) treatments without PAR were given. Chondrus crispus collected from 4–6 m depth contained only traces of the MAA palythine. After 24 h exposure to 100% ambient PAR, traces of three additional MAAs, shinorine, palythinol and palythene, were detected, and their concentrations increased strongly during a one-week exposure to all PAR treatments. The concentration of all MAAs varied directly with PAR dose, with palythine and shinorine being four- to sevenfold higher than palythinol and palythene. Likewise, naturally high doses of both UV-A and UV-B resulted in a strong accumulation of all MAAs, in particular shinorine. While shinorine accumulation was much more stimulated by UVR, the content of all other MAAs was more affected by high PAR, indicating an MAA-specific induction triggered by UVR or PAR. Received: 24 September 1997 / Accepted: 17 December 1997     

Differential responses of tetrasporophytes and gametophytes of Mazzaella laminarioides (Gigartinales,Rhodophyta) under solar UV radiation

The effects of solar UV radiation on mycosporine‐like amino acids (MAAs), growth, photosynthetic pigments (Chl a, phycobiliproteins), soluble proteins (SP), and C and N content of Mazzaella laminarioides tetrasporophytes and gametophytes were investigated. Apical segments of tetrasporophytes and gametophytes were exposed to solar radiation under three treatments (PAR [P], PAR+UVA [PA], and PAR+UVA+UVB [PAB]) during 18 d in spring 2009, Punta Arenas, Chile. Samples were taken after 2, 6, 12, and 18 d of solar radiation exposure. Most of the parameters assessed on M. laminarioides were significantly influenced by the radiation treatment, and both gametophytes and tetrasporophytes seemed to respond differently when exposed to high UV radiation. The two main effects promoted by UV radiation were: (i) higher synthesis of MAAs in gametophytes than tetrasporophytes at 2 d, and (ii) a decrease in phycoerythrin, phycocyanin, and SPs, but an increase in MAA content in tetrasporophytes at 6 and 12 d of culture. Despite some changes that were observed in biochemical parameters in both tetrasporophytes and gametophytes of M. laminarioides when exposed to UVB radiation, these changes did not promote deleterious effects that might interfere with the growth in the long term (18 d). The tolerance and resistance of M. laminarioides to higher UV irradiance were expected, as this intertidal species is exposed to variation in solar radiation, especially during low tide.     

Effects of UV radiation on five marine microphytobenthic Wadden sea diatoms,isolated from the Solthörn tidal flat (Lower Saxony,southern North Sea) – Part II: changes in carbohydrate,amino acid and fatty acid composition

Benthic diatoms inhabiting intertidal flats face highly variable environmental conditions, due to changing water levels and exposure during low tide. The present study is the second part of a more extensive study of the adaptive potential of these species in response to varying UV radiations in the Solthörn tidal flat (Lower Saxony, southern North Sea). Five isolates (Achnanthes exigua, Amphora exigua, Cocconeis peltoides, Diploneis littoralis and Navicula digitoradiata), which were found in this area in high cell numbers in summer 2008, were used in semi-continuous cultures to study the physiological effects of UV-radiation (PAR [photosynthetically active radiation], PAR+UV-A, PAR+UV-B, PAR+UV-B+UV-A). For short- and long-term exposures (6 h, 30 days), the composition of intercellular carbohydrates, amino and fatty acids were analysed in exponential-phase cultures grown at a salinity of 30 in a 12?:?12 h light?:?dark cycle at 20?°C. Although all tested species showed distinct differences in their initial carbohydrate, amino and fatty acid compositions and in their responses to the different UV treatments, general response patterns could be identified. Overall physiological responses to short- and long-term UV treatments included the accumulation of proline as well as an increase in total carbohydrates and lipids, whereas significant differences in the composition of carbohydrates, amino and fatty acids occurred after long-term exposure to the UV treatments (P < 0.05). While UV-A exposure led to higher accumulations of phenylalanine, aspartic acid and saturated fatty acids, the response to UV-B long-term exposure included increases of galactose, mannose and unsaturated fatty acids in the cells. In both UV experiments there was a noteworthy accumulation of the amino acid tryptophan in most species. The combined UV-A+UV-B experiment showed a significant (P < 0.05) increase of aspartic acid, phenylalanine, galactose and saturated fatty acids in a majority of species. Overall, the results indicated significant differences in the physiological responses of the five diatom taxa during UV exposure, which suggests species-specific acclimation strategies that may explain the growth insensitivity towards at least short-term UV.     

Migrant blackbirds,Turdus merula,have higher plasma levels of polyunsaturated fatty acids compared to residents,but not enhanced fatty acid unsaturation index

Birds have been observed to have dietary preferences for unsaturated fatty acids during migration. Polyunsaturated fatty acids (PUFAs) may increase the exercise performance of migrant birds; however, PUFAs are also peroxidation prone and might therefore incur increased costs in terms of enhanced oxidative damage in migratory individuals. To shed light on this potential constraint, we analyzed plasma fatty acid (FA) composition and estimated the unsaturation index as a proxy for susceptibility to lipid peroxidation of migrants and residents of the partially migratory common blackbird (Turdus merula) at a stopover site during autumn migration. As predicted, migrant birds had higher relative and absolute levels of PUFAs compared to resident birds. This included the strictly dietary ω‐3 PUFA α‐linolenic acid, suggesting a dietary and/or storage preference for these FAs in migrants. Interestingly, the FA unsaturation index did not differ between migrants and residents. These findings suggest a mechanism where birds alter their levels of metabolic substrate without simultaneously increasing the susceptibility of the substrate to lipid peroxidation. In summary, our results are in line with the hypothesis that increased exercise performance during migration might be constrained by oxidative stress, which is manifested in changes in the composition of key FAs to retain the unsaturation index constant despite the increased levels of peroxidizable PUFAs.     

Chemical profiling of mycosporine‐like amino acids in twenty‐three red algal species

Rhodophyta produce a variety of chemically different mycosporine‐like amino acids (MAAs), compounds that are known as some of the strongest ultraviolet (UV) absorbing molecules in nature. Accordingly, they primarily act as photoprotectants against harmful levels of solar ultraviolet radiation in the UV‐A and UV‐B range. In order to get a deeper understanding of the chemical diversity of MAAs in red algae, pure standards of eleven mycosporine‐like amino acids were isolated from three different species (Agarophyton chilense, Pyropia plicata and Champia novae‐zelandiae) using various chromatographic methods. Their structures were confirmed by nuclear magnetic resonance and mass spectrometry. Four out of the eleven MAAs are reported for the first time in algae. In addition, a new high‐performance liquid chromatography method was developed for the separation of all isolated MAAs and successfully applied for the analysis of twenty‐three red algal species of marine origin. All of them contained MAAs, the most abundant compounds were shinorine, palythine, asterina‐330 and porphyra‐334. For some samples, the direct assignment of MAAs based on their UV spectra was not possible; therefore, the target analytes were enriched by a simple concentration step, followed by liquid chromatography‐mass spectrometry analysis of the extracts. This approach enabled a deeper insight into the MAA pattern of red algae, indicating that not only the four dominant ones are synthesized but also many others, which were often described as unknown compounds in previous studies.     

The essential dietary fatty acid requirement of the tufted apple budmoth, Platynota idaeusalis

The tufted apple budmoth, Platynota idaeusalis (Walker), was reared non-axenically for two successive generations on a casein-based semisynthetic diet. The qualitative essential fatty acid requirement for growth, development and normal pupal-adult ecdysis was studied using the non-axenic casein-based semisynthetic diets with and without various 99% pure fatty acids. Linoleic or linolenic acids caused accelerated larval development; linoleic, linolenic and arachidonic acids showed similar activity in body weight gain and survival to pupal-adult ecdysis. Linoleic or linolenic acids were active in alleviating wing deformities; arachidonic acid was partially active in alleviating wing deformities at the one dietary concentration evaluated. Activity of arachidonic acid as an essential fatty acid for P. idaeusalis is unique among insects, except for mosquitoes. The essential fatty acid deficiency syndrome of the adult, resulting from the larvae feeding on fat-deficient diets, was greatly reduced when larvae were fed on a diet adequate in essential fatty acid during either their early or late development.     

Effects of artificial ultraviolet-B radiation on growth and fatty acid composition of duckweed (Lemna minor)

1. Duckweed (Lemna minor), collected either in summer or early fall was exposed under laboratory conditions to control (photosynthetically active and UV‐A radiation) or experimental (control plus UV‐B radiation) conditions. 2. Growth and survival were determined by counting the number of green, and brown/white fronds following 1–5 or 11 days of irradiation. Growth of duckweed was impaired by exposure to UV‐B radiation in the fall experiment but not in the summer. 3. Fatty acid compositions were analysed following 5 or 11 days of irradiation and a recovery period of 0, 5, 29 or 40 h. Concentrations of the major fatty acids, palmitic, linoleic (LA) and α‐linolenic (ALA) acids were similar in the summer and fall duckweed collections, but the summer samples had higher concentrations of the desaturation products of LA and ALA. 4. UV‐B exposure had small, but significant, and contrasting effects on duckweed fatty acid concentrations. In the summer experiment, duckweed exposed to UV‐B had slightly lower concentrations of major fatty acids than control duckweed, while the reverse was true in the fall experiments. 5. These minor effects of UV‐B on concentrations of LA and ALA would be unlikely to have a major impact on the supply of these essential fatty acids from duckweed to freshwater food webs.     

UV-A Mediated Modulation of Photosynthetic Efficiency,Xanthophyll Cycle and Fatty Acid Production of <Emphasis Type="Italic">Nannochloropsis</Emphasis>

Nannochloropsis, a green microalga, is a source for commercially valuable compounds as extensively described and, in particular, is recognised as a good potential source of eicosapentaenoic acid (20:5ϖ3), an important polyunsaturated fatty acid for human consumption for prevention of several diseases. Climate change might include variation in the ultraviolet (UV) levels as one of the consequences derived from the anthropogenic activity. This paper shows the response of Nannochloropsis cultures exposed for 7 days to UV-A (320–400 nm) added to photosynthetically active radiation (PAR; 400–700 nm). Growth rates and photosynthetic activity were assessed to determine the impact of UV-A increased levels on the cell growth and basic metabolism activity. Xanthophyll pigments (zeaxanthin and violaxanthin), carotenoids (canthaxanthin and β-carotene) and polyunsaturated fatty acids (myristic, palmitic, palmitoleic, arachidonic and eicosapentaenoic acids) were measured for assessing the antioxidant response of the microalgae to added UV-A radiation to PAR. The results show that the modulated use of UV-A radiations can lead to increased growth rates, which are sustained in time by an increased light transduction activity. The expected antioxidant response to the incident UV-A radiation consisted of increases in zeaxanthin and β-carotene contents—synthesis of antioxidant carotenoids—and increases in the saturated fatty acids to polyunsaturated fatty acids ratio. The results suggest that modulated UV-A radiation can be used as a tool to stimulate value molecules accumulation in microalgae through an enhanced both light transduction process and antioxidant response, while sustaining cell growth.     

Chemical Composition during Maturing and Spawning of the Sponge Dysidea herbacea (Porifera: Demospongiae)

Mycosporine-like amino acids (MAAs) have been implicated in many biochemical processes in marine organisms, but the major emphasis has been directed to their role as UV protectant compounds. The quantitation of MAAs, mycosporine-glu-gly, mycosporine-gly, usujirene and palythene in the sponge Dysidea herbacea [34] suggests that whereas some mycosporine amino acids may serve in this capacity, others are intrinsically involved in the reproductive process. The role of other compounds, such as homarine, gadusol and arachidonic acid, in reproduction of this sponge is also discussed.     

MAA Synthesis and Accumulation in Polar Macroalgae are Controlled by Abiotic Factors

Mycosporine‐like amino acids (MAAs) are regarded as powerful sunscreens protecting the algae against harmful UV radiation. The MAA protection efficiency was tested in algal samples by measuring the optimum quantum yield of photosynthesis using photosystem II fluorescence. It could be demonstrated that the recovery of photosynthesis after exposure to enhanced UV radiation is faster in individuals with high MAA content. MAAs can be synthesized in several polar macroalgae in response to different radiation conditions. Although MAA induction patterns are very species‐specific, some similarities can be found. Field studies indicate that plants from different growth habitats providing distinct radiation climate can be grouped into three physiological categories depending on their MAA content. The first group (I) includes mainly deep‐water species, typically lacking MAAs. The second group (II), algal species found in a broad range of water depths (eu‐ and sublittoral), which are able to flexibly synthesize and accumulate MAAs. The third group (III) includes supra‐ and eulittoral taxa, which always contain high MAA concentrations. In laboratory studies, we showed that taxa of group II and III responded in three different ways based on MAA accumulation when exposed to different radiation conditions (PAR, PAR + UVA, PAR + UVA + UVB). Either they: (a) exhibit highest total MAA concentration under the full artificial spectrum; (b) increase their MAA concentration after exposure to PAR and PAR + UVA or (c) MAA concentration declines after exposure to the full spectrum. Our studies have indicated that when coupled with UVR, exposure to temperature fluctuations ranging from 0 to 10 °C also affect MAA biosynthesis.     

Influence of fatty acids and bovine serum albumin on the growth of human hepatoma and immortalized human kidney epithelial cells

Summary The protective influence of bovine serum albumin against growth inhibition caused by fatty acids was studied in human hepatoma (HepG2) and immortalized human kidney epithelial (IHKE) cells. In general, growth inhibition by unsaturated fatty acids (0.15 mmol/liter) increased with increasing number of double bonds. For HepG2 cells crude albumin (1g/100 ml) did not greatly modify growth inhibition by arachidonic, eicosapentaenoic, and docosahexaenoic acid. With oleic, linoleic, and linolenic acids, crude and defatted albumin stimulated cell growth. In contrast, for IHKE cells both albumins counteracted growth inhibition by unsaturated fatty acids to approximately the same extent. When HepG2 cells were cultured in the presence of saturated fatty acids (0.3 mmol/liter), C2, C6, and C8 had no or little inhibitory effect. C10 and C12 inhibited cell growth appreciably, whereas C14, and especially C16, had poor inhibitory effects. Crude albumin counteracted growth inhibition by all these fatty acids. In contrast, defatted albumin had little or no effect (except against C10 and C12), and even increased the growth inhibition by C14 and C16. With unsaturated fatty acids there seemed to be an inverse relationship between cell growth and the concentration of thiobarbituric acid reactive substances (TBARS) in media. Vitamin E abolished growth inhibition (and the increase in TBARS concentration) by unsaturated fatty acids. The complex interaction between fatty acids and albumins calls for great caution when interpreting data on growth effects.     

阳光紫外辐射对室内水培发状念珠藻生理特性的影响

发状念珠藻（Nostoc flagelliforme Bornet & Flahault）是一种重要的陆生经济蓝藻,室内培育出的原植体如何适应阳光辐射的问题尚需探讨。为此,作者将室内水培发菜置于阳光下培养,测定了其生长、有效光化学效率（F/Fm&#900;）和色素的变化。结果表明,较高的可见光（PAR,395－700 nm）和紫外辐射（UVR,280－395 nm）均导致水培发菜的F/Fm&#900;下降。第1天中午,PAR和UVR分别使F/Fm&#900;下降了54％和13％;傍晚,F/Fm&#900;有部分恢复。UVR对发菜适应阳光2d后的生长无负面作用。发菜在适应全阳光辐射期间,紫外吸收物质（Scytonemin和Mycosporine-like amino acids）含量不断增加,9d后,分别增加了124倍和9倍。这些紫外吸收物质的增加对发菜细胞降低光抑制,适应阳光辐射,起到了重要作用。本研究的结果可为水培发菜室外培养方法的建立提供一定的理论依据。     

Serum fatty acid analysis and digestibility study in the Vancouver Island marmot (Marmota vancouverensis) fed a captive diet supplemented with α‐linolenic acid

Mammals that hibernate must rely on endogenous lipid reserves to survive over winter. This study was conducted to compare the difference in serum fatty acid composition, dietary intake, and apparent digestibility in the Vancouver Island marmot (N = 6) fed the Metro Zoo lagomorph diet supplemented with α‐linolenic acid [C(18:3) n‐3 (α‐LA)]. The experiment was designed as a 3 × 3 Latin square with three 17‐day collection periods. The test diets contained 12.16, 14.85, and 17.05% α‐LA as a percentage of fatty acids in the diet supplied through the addition of flaxseed oil (～53% α‐LA). Across treatments, dry matter intake (g/d), dry matter digestibility, apparent fat digestibility, and apparent neutral detergent fiber digestibility did not differ significantly (P > 0.05). There were no significant differences in serum α‐LA concentration between the three levels of α‐LA supplementation. However, this supplementation did elevate serum α‐LA, eicosapentaenoic acid [C20:5 (n‐3)], and docosahexaenoic acid [C22:6 (n‐3)] levels compared with feeding the basal zoo lagomorph diet (P < 0.05). Thus, supplementation of the basal zoo lagomorph diet with α‐LA elevated the serum levels of essential fatty acids in the Vancouver Island marmot. Zoo Biol 20:251–259, 2001. © 2001 Wiley‐Liss, Inc.     

Characterization of an enriched lipoxygenase extract from Aspergillus niger in terms of specificity and nature of flavor precursors production

Aspergillus niger was grown for 6 days, and the harvested biomass was homogenized; the resultant supernatant, considered as the crude enzymatic extract, was enriched by ammonium sulfate precipitation. The extract was assayed for its lipoxygenase (LOX) activity using a wide range of polyunsaturated fatty acids (PUFAs), including linoleic, linolenic and arachidonic acids, as substrates. Two pH maxima were determined at 5.0, 10.5. The K_m and V_max values indicated that the microbial LOX displayed preferential substrate specificity towards linolenic acid at low pH. The microbial LOX demonstrated preferential substrate specificity towards free fatty acids over the acyl esters of linoleic acid. It was shown that the LOX activity of A. niger produced all monohydroperoxy regioisomers of the PUFAs, and there was a predominance of conjugated diene hydroperoxides. Significant production of the unconjugated 10-hydroperoxides of both linoleic and linolenic acids was obtained by the LOX activity. The amounts of 10-hydroperoxides ranged from 15 to 21% of total produced isomers, for linolenic and linoleic acids, respectively. The greatest proportion of the 10-regioisomer was attributed to the maximum activity at pH 5.0. Four major hydroperoxy-eicosatetraenoic acid (HPETE) regioisomers were isolated from the bioconversion of arachidonic acid, including the 8-, 9-, 12- and 15-HPETE, which accounted for approximately 97% of total isomers.     

Effect of growth temperature on lipid fatty acids of four fungi (Aspergillus niger,Neurospora crassa,Penicillium chrysogenum,andTrichoderma reesei)

The effect of growth temperature on the lipid fatty acid composition was studied over a temperature range from 35 to 10° C with 5° C intervals in four exponentially growing fungi: Aspergillus niger, Neurospora crassa, Penicillium chrysogenum, and Trichoderma reesei. Fatty acid unsaturation increased in A. niger, P. chrysogenum, and T. reesei when the temperature was lowered to 20–15, 20, and 26–20° C, respectively. In A. niger and T. reesei, this was due to the increase in linolenic acid content. In P. chrysogenum, the linolenic acid content increased concomitantly with a more pronounced decrease in the less-unsaturated fatty acid, oleic acid, and in palmitic and linoleic acids; consequently, the fatty acid content decreased as the temperature was lowered to 20° C. In T. reesei, when the growth temperature was reduced below 26–20° C, fatty acid unsaturation decreased since the mycelial linolenic acid content decreased. In A. niger and P. chrysogenum, the mycelial fatty acid content increased greatly at temperatures below 20–15° C. In contrast, in N. crassa, fatty acid unsaturation was nearly temperature-independent, although palmitic and linoleic acid contents clearly decreased when the temperature was lowered between 26 and 20° C; concomitantly, the growth rate decreased. Therefore, large differences in the effects of growth temperature on mycelial fatty acids were observed among various fungal species. However, the similarities found may indicate common regulatory mechanisms causing the responses. Received: 1 March 1995 / Accepted: 8 May 1995     

Metabolism of n-3 polyunsaturated fatty acids and modification of phospholipids in cultured rabbit aortic smooth muscle cells

The metabolism of the linolenic acid family (n-3) of fatty acids, e.g., linolenic, eicosapentaenoic, and docosahexaenoic acids, in cultured smooth muscle cells from rabbit aorta was compared to the metabolism of linoleic and arachidonic acids. There was a time-dependent uptake of these fatty acids into cells for 16 hr (arachidonic greater than docosahexaenoic, linoleic, eicosapentaenoic greater than linolenic), and the acids were incorporated mainly into phospholipids and triglycerides. Eicosapentaenoic and arachidonic acids were incorporated more into phosphatidylethanolamine and phosphatidylinositol plus phosphatidylserine and less into phosphatidylcholine than linolenic and linoleic acids. Docosahexaenoic acid was incorporated into phosphatidylethanolamine more than linolenic and linoleic acids and into phosphatidylinositol plus phosphatidylserine less than eicosapentaenoic and arachidonic acids. Added linolenic acid accumulated mainly in phosphatidylcholine and did not decrease the arachidonic acid content of any phospholipid subfraction. Elongation-desaturation metabolites of linoleic acid did not accumulate. Cells treated with eicosapentaenoic acid accumulated both eicosapentaenoic and docosapentaenoic acids mainly in phosphatidylethanolamine and the arachidonic acid content was decreased. Added docosahexaenoic acid accumulated mainly in phosphatidylethanolamine and decreased the content of both arachidonic and oleic acids. The following conclusions are drawn from these results. The three n-3 fatty acids are utilized differently in phospholipids. The arachidonic acid content of phospholipids is reduced by eicosapentaenoic and docosahexaenoic acids, but not by linolenic acid. Smooth muscle cells have little or no desaturase activity, but have significant elongation activity for polyunsaturated fatty acids.